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Available from: Gary Siuzdak, Jan 16, 2014
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    • "This has prompted research into suitable alternatives to viral and plasmid reprogramming (Okita et al., 2008; Woltjen et al., 2009; Yu et al., 2009; Zhou et al., 2009; Li et al., 2011; Worsdorfer et al., 2013; Talluri et al., 2014). The SB and PB transposon systems share many advantages for mediating epigenetic reprogramming, and possess a high cargo capacity of >100 kb (Rostovskaya et al., 2012). "
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    ABSTRACT: Induced pluripotent stem cells (iPSCs) are a seminal breakthrough in stem cell research and are promising tools for advanced regenerative therapies in humans and reproductive biotechnology in farm animals. iPSCs are particularly valuable in species in which authentic embryonic stem cell (ESC) lines are yet not available. Here, we describe a nonviral method for the derivation of bovine iPSCs employing Sleeping Beauty (SB) and piggyBac (PB) transposon systems encoding different combinations of reprogramming factors, each separated by self-cleaving peptide sequences and driven by the chimeric CAGGS promoter. One bovine iPSC line (biPS-1) generated by a PB vector containing six reprogramming genes was analyzed in detail, including morphology, alkaline phosphatase expression, and typical hallmarks of pluripotency, such as expression of pluripotency markers and formation of mature teratomas in immunodeficient mice. Moreover, the biPS-1 line allowed a second round of SB transposon-mediated gene transfer. These results are promising for derivation of germ line-competent bovine iPSCs and will facilitate genetic modification of the bovine genome.
    Cellular Reprogramming 04/2015; DOI:10.1089/cell.2014.0080. · 2.35 Impact Factor
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    • "Even the seemingly straightforward DNA-free non-integrating methods [e.g. small molecule cocktails (Hou et al, 2013) or proteins (Zhou et al, 2009)] remain highly inefficient, requiring intricate protocols with multiple rounds of treatment, large doses, and uncontrolled presence/kinetics of reprogramming factors. Thus, new strategies are required to overcome the limitations associated with these traditional reprogramming methods. "
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    ABSTRACT: Derived from any somatic cell type and possessing unlimited self-renewal and differentiation potential, induced pluripotent stem cells (iPSCs) are poised to revolutionize stem cell biology and regenerative medicine research, bringing unprecedented opportunities for treating debilitating human diseases. To overcome the limitations associated with safety, efficiency, and scalability of traditional iPSC derivation, expansion, and differentiation protocols, biomaterials have recently been considered. Beyond addressing these limitations, the integration of biomaterials with existing iPSC culture platforms could offer additional opportunities to better probe the biology and control the behavior of iPSCs or their progeny in vitro and in vivo. Herein, we discuss the impact of biomaterials on the iPSC field, from derivation to tissue regeneration and modeling. Although still exploratory, we envision the emerging combination of biomaterials and iPSCs will be critical in the successful application of iPSCs and their progeny for research and clinical translation. © 2015 The Authors.
    The EMBO Journal 03/2015; 34(8). DOI:10.15252/embj.201490756 · 10.75 Impact Factor
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    • "To decrease tumorigenic potential different methods of generating hiPSCs were developed, including different combinations of reprogramming genes [22] [23] together with small molecules [24], which increased reprogramming efficiency, and use of different excisable [11] [25] [26] or nonintegrative vectors [27] for delivering reprogramming factors. A step forward to virus-free reprogramming methods was application of synthetic-modified mRNA [28] and recombinant proteins [29] [30] [31]. Therefore, application of improved reprogramming protocols for hiPSC generation, which will be safe for clinical use and production of patient-specific iPSC-derived cardiomyocytes (iPSC-CMs), would theoretically overcome immunological complications of transplanting organs and enable avoiding immunosuppressive treat- ment. "
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    ABSTRACT: Human pluripotent stem cells (hPSCs), namely, embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), with their ability of indefinite self-renewal and capability to differentiate into cell types derivatives of all three germ layers, represent a powerful research tool in developmental biology, for drug screening, disease modelling, and potentially cell replacement therapy. Efficient differentiation protocols that would result in the cell type of our interest are needed for maximal exploitation of these cells. In the present work, we aim at focusing on the protocols for differentiation of hPSCs into functional cardiomyocytes in vitro as well as achievements in the heart disease modelling and drug testing on the patient-specific iPSC-derived cardiomyocytes (iPSC-CMs).
    04/2014; 2014:512831. DOI:10.1155/2014/512831
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